Gelsolin was labelled with the sulphydryl-specific fluorescent reagent 6-acryloyl-2-dimethylaminonaphthalene (acrylodan). The degree of labelling using non-denaturing conditions was 1.9 ± 0.5 acrylodans per gelsolin molecule. Circular dichroism and viscosity studies showed no significant effect on gelsolin structure and function on incorporation of the label.
Circular dichroism studies did not detect Ca²⁺ effects on aerylodan-labelled gelsolin, but fluorescence studies detected subtle changes in the protein. The presence of Ca²⁺ causes a decrease and red-shift in fluorescence emission, an increase in sensitivity to quenching by I⁻ and a decrease in fluorescence polarisation of the acrylodan-labelled gelsolin. These indicate an increased degree of exposure of the fluorescent label to the solvent environment on interaction of gelsolin with Ca²⁺.
Actin binding to gelsolin was evident from a decrease in fluorescence intensity, an increase in sensitivity to quenching and an increase in fluorescence polarisation. Actin binding increases the exposure of the acrylodan label to solvent, as does Ca²⁺ binding. / Science, Faculty of / Chemistry, Department of / Graduate
| Identifer | oai:union.ndltd.org:UBC/oai:circle.library.ubc.ca:2429/30278 |
| Date | January 1990 |
| Creators | Reid, Scott William |
| Publisher | University of British Columbia |
| Source Sets | University of British Columbia |
| Language | English |
| Detected Language | English |
| Type | Text, Thesis/Dissertation |
| Rights | For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use. |
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